李婷; 伍平; 陈曼; 王爱先; 傅旻婧; 吴雪英; 甄军毅; 宫美维; 王卉

doi:10.3760/cma.j.cn114452-20240312-00122

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多参数流式细胞术筛选急性早幼粒细胞白血病免疫表型特征

中华检验医学杂志, 2024,47(7) : 747-754. DOI: 10.3760/cma.j.cn114452-20240312-00122

摘要

目的

多参数流式细胞术（MFC）分析筛选急性早幼粒细胞白血病（APL）免疫表型特征。

方法

回顾性描述性研究。纳入2016年1月1日至2023年12月31日于河北燕达陆道培医院就诊的130例急性髓系白血病（AML）患者，其中经典型APL（cAPL）组44例，微颗粒变异型APL（APLv）组24例，非APL［包括NPM1突变AML（NPM1^mut AML）、AML伴KMT2A重排等］组62例。采用MFC免疫分型法分析和比较各组侧向散射光（SSC）中位表达强度（MEI）及其白血病细胞与自身成熟淋巴细胞MEI比值（T/L MEIR），CD34、髓过氧化物酶（MPO）、CD64和CD9的中位荧光强度（MDFI）及其上述参数白血病细胞与自身成熟淋巴细胞MDFI比值（T/L MDFIR）差异；绘制受试者工作特征（ROC）曲线评价流式多参数模型对cAPL和非APL、APLv和非APL的鉴别诊断效能。

结果

cAPL组SSC的MEI、T/L MEIR高于APLv组和非APL组（P均<0.05），且APLv组高于非APL组（P<0.05）；cAPL组和APLv组CD34的MDFI均高于非APL组（P均<0.05），APLv组的CD34 T/L MDFIR高于非APL组（P<0.05）；cAPL组和APLv组MPO、CD9的MDFI、T/L MDFIR均高于非APL组（P均<0.05）；cAPL组CD64的MDFI、T/L MDFIR均高于非APL组（P均<0.05）。ROC曲线结果显示，SSC的MEI、CD64的MDFI、CD9的MDFI、SSC的T/L MEIR和CD9的T/L MDFIR诊断cAPL的ROC曲线下面积（AUC）分别为0.932、0.816、0.893、0.960和0.894，诊断APLv的AUC分别为0.725、0.737、0.791、0.729和0.736，其对鉴别cAPL和非APL、APLv和非APL均有较好的诊断价值（P均<0.05）。

结论

通过MFC方法可分析筛选APL的免疫表型特征，对cAPL、APLv和非APL患者进行鉴别诊断。

引用本文: 李婷, 伍平, 陈曼, 等. 多参数流式细胞术筛选急性早幼粒细胞白血病免疫表型特征 [J] . 中华检验医学杂志, 2024, 47(7) : 747-754. DOI: 10.3760/cma.j.cn114452-20240312-00122.

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急性早幼粒细胞白血病（acute promyelocytic leukemia，APL）伴PML：：RARA融合基因占新诊断的急性髓系白血病（acute myeloid leukemia，AML）患者的10%~15%^［1］。APL病程进展迅速，如不能及时治疗，患者病死率高^［2］，因此快速诊断和及时治疗至关重要^［3］。APL根据形态学可分为2种类型：经典型APL（classical APL，cAPL）（即多颗粒型APL）和微颗粒变异型APL（APL，hypogranular variant，APLv），典型免疫表型为表达分化抗原簇（cluster of differentiation，CD）13、CD33、CD117和髓过氧化物酶（myeloperoxidase，MPO）和CD64，不表达人类白细胞抗原DR（human leukocyte antigen DR，HLA-DR）、CD34、CD11b和CD14。然而，AML中的非APL［例如核磷蛋白1突变（nucleophosmin 1 mutation，NPM1^mut）AML和伴有赖氨酸甲基转移酶2A（lysine methyltransferase 2A，KMT2A）重排的AML］，也可表现出类似APLv的细胞形态特征和免疫表型^{［4, 5］}。目前国内外主要通过多参数流式细胞术（multiparameter flow cytometry，MFC）检测细胞表面抗原（如CD64、CD117、CD11b、CD34、CD33、CD13和HLA-DR）的表达情况鉴别APL和非APL^{［6, 7］}。本研究通过分析具有APL样免疫表型的AML（包括cAPL，APLv和非APL）患者的免疫表型谱，评估MFC免疫表型分析在cAPL、APLv和非APL鉴别诊断中的应用价值。

贡献者信息

李婷

北京陆道培医院病理与医学检验科，北京　100176

伍平

河北燕达陆道培医院检验科，廊坊　065201

陈曼

河北燕达陆道培医院检验科，廊坊　065201

王爱先

河北燕达陆道培医院检验科，廊坊　065201

傅旻婧

北京陆道培医院病理与医学检验科，北京　100176

吴雪英

河北燕达陆道培医院检验科，廊坊　065201

甄军毅

河北燕达陆道培医院检验科，廊坊　065201

宫美维

河北燕达陆道培医院检验科，廊坊　065201

王卉

河北燕达陆道培医院检验科，廊坊　065201

通信作者

王卉

河北燕达陆道培医院检验科，廊坊　065201

Email：wh9784@163.com

关键词

白血病，早幼粒细胞，急性; 流式细胞术; 免疫表型分型; 中位荧光强度; 鉴别诊断;

作者声明

作者贡献声明

李婷：参与设计实验、采集数据、分析/解释数据、统计分析和文章撰写；陈曼、甄军毅、宫美维：参与设计实验、采集数据；伍平、王爱先、傅旻婧、吴雪英、王卉：参与分析/解释数据、统计分析和文章撰写；王卉：行政、技术或材料支持和指导工作

利益冲突

所有作者声明无利益冲突

历史

出版日期：2024-07-19

收稿日期：2024-03-12

本文编辑

干岭

Original Article

Screening the immunophenotypic characteristics of acute promyelocytic leukemia by multiparameter flow cytometry

Li Ting, Wu Ping, Chen Man, Wang Aixian, Fu Minjing, Wu Xueying, Zhen Junyi, Gong Meiwei, Wang Hui

Published 2024-07-19

Cite as Chin J Lab Med, 2024, 47(7): 747-754. DOI: 10.3760/cma.j.cn114452-20240312-00122

Abstract

Objective

To explore and screen the immunophenotypic characteristics of acute promyelocytic leukemia (APL) by multiparameter flow cytometry (MFC).

Methods

A retrospective and descriptive study. A total of 130 acute myeloid leukemia (AML) patients who registrated in Hebei Yanda Lu Daopei Hospital were studied, among which there were 44 classical APL (cAPL), 24 microgranular variant of APL (APLv) and 62 non-APL patients (including NPM1^mut AML and AML with KMT2A rearrangement). MFC immunotyping was used to analyze and compare the median expression intensity (MEI) of side scatter (SSC), along with the ratio of the MEI on leukemic cells with those on lymphocytes (T/L MEIR), the median fluorescence intensity (MDFI) of CD34, myeloperoxidase (MPO), CD64 and CD9 on leukemic cells, as well as the ratios of these MDFIs on leukemic cells with those on lymphocytes (T/L MDFIR). Receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic efficiency of the multiparameters model for distinguishing cAPL and non-APL, APLv and non-APL.

Results

The MEI and T/L MEIR of SSC in the cAPL group were higher than those in the APLv and non-APL groups (P<0.05), and these two parameters in APLv group were higher than those in the non-APL group, respectively (P<0.05). The MDFIs of CD34 in cAPL and APLv groups were higher than those in the non-APL group (P<0.05), and the T/L MDFIR of CD34 was higher in APLv group than non-APL group (P<0.05). The MDFIs of MPO and CD9, as well as the T/L MDFIRs in cAPL and APLv groups were both higher than those in the non-APL group, respectively (P<0.05). The MDFI and T/L MDFIR of CD64 in the cAPL group were higher than those in non-APL group, respectively (P<0.05). ROC curve results showed that the area under the curve (AUC) of MEI of SSC, the MDFI of CD64 and CD9, as well as the T/L MEIR of SSC and T/L MDFIR of CD9 were 0.932, 0.816, 0.893, 0.960 and 0.894 for diagnosing cAPL, respectively, and the AUC of these parameters were 0.725, 0.737, 0.791, 0.729 and 0.736 for diagnosis APLv, respectively (P<0.05).

Conclusion

MFC method can analyze and screen the immunophenotypic characteristics of APL for differential diagnosis of cAPL, APLv and non-APL patients.

Key words:

Leukemia, promyelocytic, acute; Flow cytometry; Immunophenotyping; Median fluorescence intensity; Differential diagnosis

Contributor Information

Li Ting

Department of Pathology and Medical Laboratory, Beijing Lu Daopei Hospital, Beijing 100176, China

Wu Ping